US4476225A - Apparatus for continuously disintegrating cells of microorganisms - Google Patents

Apparatus for continuously disintegrating cells of microorganisms Download PDF

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Publication number
US4476225A
US4476225A US06/397,122 US39712282A US4476225A US 4476225 A US4476225 A US 4476225A US 39712282 A US39712282 A US 39712282A US 4476225 A US4476225 A US 4476225A
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US
United States
Prior art keywords
sleeve
suspension
valve member
mixing vessel
cells
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US06/397,122
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English (en)
Inventor
Alfred N. Grigorian
Andrei P. Kovalev
Vitaly V. Lalov
Nikolai D. Makarov
Rady V. Katrush
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VSESOJUZNY NAUCHNO-ISSLEDOVATELSKY INSTITUT SINTEZA BELKOV
Vsesojuzny Nauchnoissledovatelsky Institut Sinteza Belkov
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Vsesojuzny Nauchnoissledovatelsky Institut Sinteza Belkov
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Assigned to VSESOJUZNY NAUCHNO-ISSLEDOVATELSKY INSTITUT SINTEZA BELKOV reassignment VSESOJUZNY NAUCHNO-ISSLEDOVATELSKY INSTITUT SINTEZA BELKOV ASSIGNMENT OF ASSIGNORS INTEREST. Assignors: GRIGORIAN, ALFRED N., KATRUSH, RADY V., KOVALEV, ANDREI P., LALOV, VITALY V., MAKAROV, NIKOLAI D.
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/06—Lysis of microorganisms
    • C12N1/066—Lysis of microorganisms by physical methods
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material

Definitions

  • the invention relates to the microbiological industry, and more particularly, to an apparatus for continuously disintegrating cells of microorganisms such as yeast, bacteria, water plants and fungi for isolating intracellular proteins, nucleic acids, enzymes and other physiologically active substances.
  • microorganisms such as yeast, bacteria, water plants and fungi for isolating intracellular proteins, nucleic acids, enzymes and other physiologically active substances.
  • extrusion apparatus ballistical disintegrators and apparatus using the effect of decompression, wherein the cells are destructed on the account of expansion of gas dissolved in the cells under pressure upon an abrupt pressure decrease in suspension.
  • the invention relates to the latter type of apparatus.
  • Prior are apparatus for decompression disintegration consist of two basic units.
  • the first unit comprises one or two high-pressure vessels having pipelines for feeding thereto a cell suspension and a working gas (nitrogen, air, carbon dioxide and the like) under pressure for saturating the suspension.
  • the vessels can have a suspension level regulator and stirring devices.
  • the second basic unit is a disintegration device which comprises a specially designed valve ensuring an abrupt pressure decrease in the flow of the cell suspension.
  • the prior art apparatus for decompression disintegration of cells of microorganisms differ from one another mainly in the construction of the disintegration device.
  • the disintegration device comprises a construction consisting of a top flange casing and a bottom flange casing which are vertically movable and have disk seats installed therein, the disk seat of the bottom casing having an annular projection defining with the outer surface of the top seat a working aperture, and the top casing has a passage for admitting the suspension to the working aperture (cf. USSR Inventor's Certificate No. 602550, Int. Cl. C 12 K 1/10).
  • a similar construction of the disintegration device is used, which differs in that the seat of the bottom casing has an additional annular projection and a passage for the removal of the suspension of disintegrated cells, the disk seat of the top casing having a number of grooves communicating with the passage admitting the suspension for distributing the suspension into the working aperture; in addition, an agitator is provided on the top casing for cleaning the working aperture upon clogging (cf. USSR Inventor's Certificate No. 602551, Int. Cl. C 12 K 1/10).
  • a cell suspension and a compressed gas are continuously fed to a mixing vessel having a float-type level gauge.
  • the level gauge opens the admission of compressed gas thereto, the gas bubbling under pressure through the suspension layer. Further, the suspension saturated with the gas passes through a receiver and is admitted to the disintegration device.
  • the suspension is fed to the disk seat of the top casing and is then throttled through the aperture defined between the outer surface of the top seat and the annular projection (or projections) of the bottom seat to pass to the zone of normal pressure.
  • the cells are thus disintegrated under the action of a decompression.
  • the working pressure in the disintegration device is set-up by tightening adjusting screws and is measured by means of a pressure gauge.
  • the apparatus for decompression disintegration are characterized by a high enough efficiency of breaking of the cell membranes with metal mass and energy consumption which are much lower than in apparatus of other types.
  • the apparatus function at a comparatively low working pressure (between 80 and 200 atm.g.).
  • the decompression method used in such apparatus makes it possible to scale-up such apparatus to the capacity required in the commercial production.
  • the abovedescribed apparatus are, however, deficient in a complicated construction of the disintegration device and the need in manual adjustment of pressure in the disintegration device during operation.
  • An apparatus for continuously disintegrating cells of microorganisms disclosed in USSR Inventor's Certificate No. 492118, Int. Cl. C 12 1/10 comprises a mixing vessel for allowing a suspension of microorganisms to stay under pressure, a pipeline of a disintegration device connected thereto and to an accumulating vessel for disintegrated material. Pipelines connected to a compressed gas source and to a metering pump for supplying the suspension are connected to the mixing vessel containing the suspension.
  • the apparatus has an automatic valve for maintaining a pre-set pressure in the vessel.
  • the disintegration device comprises two casings connected in series to one another and defining a supply chamber and a working chamber.
  • the supply chamber has an inlet passage for feeding the compressed gas and an outlet chamber for pressure decrease, and the working chamber has a lateral inlet passage for admitting the suspension of cells; the chambers are separated by a membrane to which is secured a valve member installed in the working chamber.
  • the casing of the working chamber is connected in series with an adjusting flange body and a needle seat body, the needle bearing with its working end against the seat, so that the seat aperture is closed on the side of the high pressure zone.
  • the apparatus functions in the following manner.
  • a working pressure in the supply chamber of the disintegration device is built-up by the compressed gas.
  • the suspension of cells is fed to the mixing vessel by the metering pump.
  • the pre-set working pressure about 100 atm.g.
  • the compressed gas is admitted to the vessel, and the suspension saturated with the gas is discharged through the distintegration device into the accumulating vessel.
  • the uniformity of decompression of the cell suspension is ensured by means of the adjusting flange which is bolted to the casing of the working chamber of the disintegration device.
  • the apparatus functions continuously and enables the degree of disintegration of cells of microorganisms at a rate of 70-75%.
  • the arrangement of the needle valve inside the working chamber and the lateral admission of the suspension to the working chamber in the abovedescribed apparatus result in swirling of the suspension flow in the passage defined between the working chamber walls and the needle and also in swirling of the suspension jet discharged through the aperture defined by the needle and its seat.
  • the time of pressure decrease in the suspension increases thus lowering the efficiency of disintegration.
  • the movement of the suspension jet at the outlet from the disintegration device determined by the shape of the aperture between the needle and its seat is directed toward the needle axis thus impairing the atomization of the suspension, hence, lowering the efficiency of disintegration.
  • the construction of the disintegration device consisting of four series-connected elements--the supply chamber, the working chamber, the adjusting flange and the needle seat--is difficult to adjust upon a change in the operating conditions and replacement of various components, thus making the apparatus as a whole more complicated in structure.
  • extrusion disintegration apparatus the cells are disintegrated by throttling the cell suspension from a high-pressure zone through a microaperture of a disintegration valve into a normal pressure zone.
  • One of the most popular apparatus of this type is a hydraulic extruder manufactured by the U.S. Company Gaulin (cf. Rees, L. H., Chem. Engineering, 1974, 5, 13, 87).
  • the apparatus structurally consists of two basic units: a high-capacity plunger pump and a spring-biased cuneiform disintegrating valve which is connected to the pump discharge. Both units are installed on a cast iron bed, together with a motor.
  • the hydraulic pressure at the disintegration valve is adjusted either manually, by means of a handwheel, or remotely.
  • the apparatus is designed for operation under recirculation of the cell suspension.
  • a suspension of microorganisms is continuously admitted at a high pressure (between 250 and 700 atm.g.) to the interior of the valve, at a low velocity.
  • the suspension then gets into a microaperture between the valve and its seat where the velocity rapidly increases, depending on the working pressure, and the suspension is throttled into the normal pressure zone where the cells of microorganisms are disintegrated to the extent determined by the value of the hydrostatic pressure, its gradient and rate of change, cavitation and turbulization effects.
  • the residence time of the cell suspension within the microaperture about 1 ⁇ m wide is 10 -6 s.
  • the process of destruction of cells in the extrusion homogeneization apparatus features high working pressure (250-700 atm.g.), and the working cycle is to be repeated many a time for achieving high degree of cell disintegration (80-90%) so that fragments of cells of different size are obtained, and the separation of the solid and liquid phases of the resultant suspension becomes very difficult.
  • an apparatus for continuously disintegrating cells of microorganisms comprising a mixing vessel for saturating cells of microorganisms in suspension with a gas under pressure, having a metering pump and pipelines for admitting a suspension and a compressed gas, respectively, and an accumulating vessel for disintegrated material which communicates with the mixing vessel through a disintegrating device comprising a hollow body having a seat in which is installed a needle valve member, according to the invention, the hollow body comprises a sleeve having a bottom wall incorporating the needle valve member and a side wall having openings for communication of the interior of the sleeve with the accumulating vessel, an insert being installed in the open end of the sleeve which is connected to the sleeve and has a central passage coaxial with the valve member and communicating with the mixing vessel, the seat in the form of a throttle washer being installed in the central passage.
  • This construction of the disintegration device makes it possible to direct the flow of suspension passing through the aperture between the needle valve member and the throttle washer in the form of a radially fanning flow to a low-pressure zone without swirling of the flow since the shut-off end of the valve member faces the orifice of the throttle washer on the low-pressure zone side. All this contributes to a high degree of disintegration of cells in suspension.
  • the needle valve member is adjustably mounted in the bottom wall of the sleeve so as to set-up a desired size of the aperture between the needle valve member and the throttle washer when the operating conditions of the apparatus in terms of pressure and/or flow rate of suspension passing through the aperture are to be changed.
  • the insert is preferably connected to the sleeve by means of a threaded joint. This facility provides for an additional adjustment of the aperture between the needle valve member and throttle washer and also facilitates assembly and disassembly of the disintegration device for replacement of the worn needle valve member.
  • the central passage of the insert diverging on the side of its entry in the throttle washer so as to provide for a free, unobstructed emergence of the suspension from the aperture between the needle valve member and throttle washer into the accumulating vessel thereby creating a higher pressure gradient in the process of disintegration and increasing the quantity of cells destructed in the disintegrator.
  • the throttle washer is preferably secured in the insert by means of a bushing and a flange which is fixed to the insert so as to ensure the alignment of the throttle washer in the insert and a rapid replacement of the throttle washer when it is worn or when the operating conditions of the disintegration device are to be changed.
  • FIG. 1 is schematic view of an apparatus, according to the invention.
  • FIG. 2 is a longitudinal sectional view of a disintegration device of the apparatus shown in FIG. 1;
  • FIG. 3 is an embodiment of a needle valve member.
  • An apparatus for continuously disintegrating cells of microorganisms comprises a mixing vessel 1 (FIG. 1) for saturating a suspension of cells of microorganisms with a gas under pressure, an accumulating vessel 2 for disintegrated material, a disintegration device 3, a pipeline 4 for communication of the mixing vessel 1 with the disintegration device 3.
  • the mixing vessel 1 is provided with a level sensor for monitoring the level of suspension in the vessel (not shown in FIG. 1), a pressure gauge 5 for measuring the pressure, pipelines 6 and 7 for admitting a suspension and a compressed gas, respectively, and a metering pump 8 communicating with the mixing vessel 1 through a pipeline 9.
  • the mixing vessel 1 may have baffles, stirrers, a heat exchanger for thermostabilization (not shown in the drawings) and other means known per se for enabling normal operation of the apparatus.
  • the disintegration device 3 is accommodated within the accumulating vessel 2 so that the mixing vessel 1 communicates with the accumulating vessel 2 through the disintegration device 3.
  • the disintegration device 3 comprises a hollow body in the form of a sleeve 10, a needle valve member 11 in the form of a needle, and a seat in the form of a throttle washer 12, the pointed end of the valve member 11 being received in the orifice of the throttle washer.
  • the other end of the needle valve member 11 is located in a low-pressure zone and is installed in a bottom wall 13 of the sleeve 10.
  • a side wall 14 of the sleeve 10 has openings 15 for communicating the interior 16 of the sleeve 10 (FIG. 1) with the accumulating vessel 2.
  • An insert 17 which is installed in the open end of the sleeve 10 (FIG. 2) is connected to the sleeve 10 and has a central passage 18 which is coaxial with the needle valve member 11 and communicates with the mixing vessel 1 through the pipeline 3 and with the accumulating vessel 2 through the interior 16, the throttle washer 12 being installed in the central passage 18.
  • the insert 17 is connected to the sleeve 10 by means of a threaded joint so as to provide for an adjustment of the position of the insert 17, together with the throttle washer 12, with respect to the valve member 11.
  • the central passage 18 of the insert 17 is made diverging on the side where the needle valve member 11 enters the throttle washer 12, as shown in FIG. 2.
  • the needle valve member 11 is installed with the possibility of adjustment in the bottom wall 13 of the sleeve 10, e.g., by means of a threaded joint defined by the bottom wall 13 of the sleeve 10 and a union nut 19 against which bears a shank of the valve member 11 which has a shoulder 20 limiting the vertical displacement of the valve member 11.
  • the union nut 19 By turning the union nut 19, the valve member 11 can be moved to adjust its position with respect to the seat, i.e. the washer 12 without disassembling the device.
  • the throttle washer 12 is secured in the insert 17 by means of a bushing 21 which is installed in the passage 18 of the insert and which is held in this position by a flange 22 of the pipeline 4, the flange 22 being fixed to the insert 17 by means of locking studs 23.
  • the needle valve member 11 may be in the form of a needle as shown in FIG. 2 or in the form of a rod 24 and a ball 25 as shown in FIG. 3.
  • the apparatus functions in the following manner.
  • a suspension of cells of microorganisms is fed to the mixing vessel 1 (FIG. 1) by means of the metering pump 8 through the pipelines 6 and 9; a compressed gas (nitrogen, air, carbon dioxide and the like) is also fed to the mixing vessel so as to bubble it through the suspension layer and to saturate the cells of microorganisms with the gas under pressure.
  • the level of suspension in the mixing vessel 1 is pre-set and then maintained constant by means of a level sensor and a level regulator (not shown in the drawings) so that the ratio of the space occupied by the gas to the volume of suspension should be between 1:3 and 3:4.
  • the working pressure is between 80 and 200 atm.g.
  • valve member 11 (FIG. 2) of the disintegration device 3 is moved vertically to open the orifice of the throttle washer 12.
  • the reciprocations of the valve member 11 may be effected either manually or automatically, by means of an actuator.
  • the suspension of microorganisms saturated with the gas starts flowing from the mixing vessel 1 through the pipeline 4 to the aperture defined between the valve member 11 and the throttle washer 12.
  • the suspension pressure decreases at a high rate in the resultant aperture to the atmospheric pressure in the accumulating vessel 2 thus causing disintegration of cells of microorganisms owing the effect of decompression.
  • the apparatus then functions continuously to reliably ensure disintegration of cells at a rate of 75-80% within predetermined ranges of suspension velocity and flow rate.
  • the employment of the apparatus according to the invention ensures an improvement of the disintegration efficiency by 5-8% owing to improved conditions for the discharge of the cell suspension jet through the throttling aperture of the disintegration device.
  • the simplification of the apparatus construction compared with the prior art makes it possible to facilitate the adjustment of the disintegration device, replacement of the valve member and throttle washer as they become worn and to automate the disintegration process.

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Cell Biology (AREA)
  • Sustainable Development (AREA)
  • Mycology (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Disintegrating Or Milling (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Crystals, And After-Treatments Of Crystals (AREA)
US06/397,122 1981-11-13 1982-07-12 Apparatus for continuously disintegrating cells of microorganisms Expired - Fee Related US4476225A (en)

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SU813339317A SU1116054A1 (ru) 1981-11-13 1981-11-13 Установка дл дезинтеграции клеток микроорганизмов
SU3339317 1981-11-13

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JP (1) JPS5889177A (US07122603-20061017-C00145.png)
CH (1) CH655514B (US07122603-20061017-C00145.png)
CS (1) CS245736B1 (US07122603-20061017-C00145.png)
DE (1) DE3226016C2 (US07122603-20061017-C00145.png)
FR (1) FR2516538B1 (US07122603-20061017-C00145.png)
SU (1) SU1116054A1 (US07122603-20061017-C00145.png)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4774187A (en) * 1986-04-14 1988-09-27 S Diessel Gmbh & Co. Cell-culture apparatus
US5308757A (en) * 1990-08-08 1994-05-03 Hitachi, Ltd. Biological cell treatment method and apparatus
US5374522A (en) * 1986-03-20 1994-12-20 Gen-Probe Incorporated Method for releasing RNA and DNA from cells
US5380826A (en) * 1989-07-20 1995-01-10 Aphios Corporation Supercritical fluid disruption of and extraction from microbial cells
US5620730A (en) * 1993-09-09 1997-04-15 Van Noort; Gerard Method of enhancing shelf-stability of an edible biological product
US7033813B2 (en) 2001-09-05 2006-04-25 Trevor P Castor Inactivated vaccines for aids and other infectious diseases
US7527734B1 (en) * 2005-11-15 2009-05-05 Shepherd Samuel L Rapid non-equilibrium decompression of microorganism-containing waste streams
US20090236270A1 (en) * 2008-03-20 2009-09-24 Industrial Technology Research Institute Water purification system
US20160199831A1 (en) * 2013-08-02 2016-07-14 All Cell Recovery LLC Systems and methods for recovering blood cells, in a controlled environment, for storage

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE4241154C1 (de) * 1992-12-07 1994-03-17 Lancaster Group Ag Verfahren zum Aufschluß von Zelldispersionen oder Zellsuspensionen mittels Ultraschallbehandlung zwecks Gewinnung von Zellinhaltsstoffen
FR2939445B1 (fr) * 2008-12-10 2013-05-03 Biomerieux Sa Systeme automatise de lyse de microorganismes presents dans un echantillon, d'extraction et de purification des acides nucleiques desdits microorganismes aux fins d'analyse.

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US3165266A (en) * 1962-02-07 1965-01-12 Sorvall Inc Ivan Cell fractionator apparatus and method
SU492118A1 (ru) * 1973-01-12 1978-04-15 Всесоюзный научно-исследовательский институт биосинтеза белковых веществ Установка дл дезинтеграции микроорганизмов
SU602551A1 (ru) * 1975-12-03 1978-04-15 Всесоюзный научно-исследовательский институт биосинтеза белковых веществ Установка дл непрерывной дезинтеграции клеток микроорганизмов
SU602550A1 (ru) * 1974-12-30 1978-04-15 Всесоюзный научно-исследовательский институт биосинтеза белковых веществ Установка дл непрерывной дезинтеграции клеток миктоорганизмов
US4084757A (en) * 1976-08-05 1978-04-18 Vladimir Jurievich Rakitin Apparatus for continuous disintegration of cells of microorganisms

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FR532555A (fr) * 1921-03-22 1922-02-07 Robinet pour brûleurs d'appareils à gaz et autres applications
GB668055A (en) * 1950-08-08 1952-03-12 Flexibox Ltd Improvements relating to stop valves
SE334212B (US07122603-20061017-C00145.png) * 1965-09-17 1971-04-19 Biox Ab
DE1958962A1 (de) * 1969-11-24 1971-05-27 Niro Atomizer As Regelventil
JPS5328989B2 (US07122603-20061017-C00145.png) * 1974-05-27 1978-08-17
FR2361466A1 (fr) * 1976-08-13 1978-03-10 Vnii Biosinteza Belkovykh Procede de desintegration en continu de cellules de micro-organismes et unite pour la mise en oeuvre dudit procede

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Publication number Priority date Publication date Assignee Title
US3165266A (en) * 1962-02-07 1965-01-12 Sorvall Inc Ivan Cell fractionator apparatus and method
SU492118A1 (ru) * 1973-01-12 1978-04-15 Всесоюзный научно-исследовательский институт биосинтеза белковых веществ Установка дл дезинтеграции микроорганизмов
SU602550A1 (ru) * 1974-12-30 1978-04-15 Всесоюзный научно-исследовательский институт биосинтеза белковых веществ Установка дл непрерывной дезинтеграции клеток миктоорганизмов
SU602551A1 (ru) * 1975-12-03 1978-04-15 Всесоюзный научно-исследовательский институт биосинтеза белковых веществ Установка дл непрерывной дезинтеграции клеток микроорганизмов
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Rees, Evaluating Homogenizers for Chemical Processing , Chemical Engineering, May 13, 1974, pp. 87 92. *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5374522A (en) * 1986-03-20 1994-12-20 Gen-Probe Incorporated Method for releasing RNA and DNA from cells
US4774187A (en) * 1986-04-14 1988-09-27 S Diessel Gmbh & Co. Cell-culture apparatus
US5380826A (en) * 1989-07-20 1995-01-10 Aphios Corporation Supercritical fluid disruption of and extraction from microbial cells
US5308757A (en) * 1990-08-08 1994-05-03 Hitachi, Ltd. Biological cell treatment method and apparatus
US5620730A (en) * 1993-09-09 1997-04-15 Van Noort; Gerard Method of enhancing shelf-stability of an edible biological product
US7033813B2 (en) 2001-09-05 2006-04-25 Trevor P Castor Inactivated vaccines for aids and other infectious diseases
US7527734B1 (en) * 2005-11-15 2009-05-05 Shepherd Samuel L Rapid non-equilibrium decompression of microorganism-containing waste streams
US20090236270A1 (en) * 2008-03-20 2009-09-24 Industrial Technology Research Institute Water purification system
US7959802B2 (en) * 2008-03-20 2011-06-14 Industrial Technology Research Institute Water purification system
US20160199831A1 (en) * 2013-08-02 2016-07-14 All Cell Recovery LLC Systems and methods for recovering blood cells, in a controlled environment, for storage
US10159980B2 (en) * 2013-08-02 2018-12-25 All Cell Recovery LLC Systems and methods for recovering blood cells, in a controlled environment, for storage

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FR2516538B1 (fr) 1986-06-20
CS245736B1 (en) 1986-10-16
FR2516538A1 (fr) 1983-05-20
JPS5889177A (ja) 1983-05-27
DE3226016A1 (de) 1983-05-19
CH655514B (US07122603-20061017-C00145.png) 1986-04-30
DE3226016C2 (de) 1986-10-02
SU1116054A1 (ru) 1984-09-30
JPS627831B2 (US07122603-20061017-C00145.png) 1987-02-19

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